SpecificityThis antibody detects a band representing Geminin by Western blot at 33kD on 293T cell lysate, this band can be blocked by the immunising peptide. It also detects a band at about 45kD that is not blocked by the immunising peptide and is thought to be non-specific.
This antibody gave a positive signal in Hek293 whole cell lysate.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500. Detects a band of approximately 33 kDa (predicted molecular weight: 23.5 kDa).
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
FunctionInhibits DNA replication by preventing the incorporation of MCM complex into prereplication complex (pre-RC). It is degraded during the mitotic phase of the cell cycle. Its destruction at the metaphase-anaphase transition permits replication in the succeeding cell cycle.
Sequence similaritiesBelongs to the geminin family.
Developmental stageAbsent during G1 phase, accumulates during S, G2, and M phases, and disappears at the time of the metaphase-anaphase transition.
The lower band represents Geminin (33 kDa - predicted molecular weight 23 kDa), and is blocked by the immunising peptide, the upper band is likely to be non-specific as it is not blocked by the peptide.
Immunocytochemistry/ Immunofluorescence - Geminin antibody (ab12147)This image is courtesy of an Abreview submitted by Dr Kirk McManus
ab12147 (1/400) staining geminin in assynchronous HeLa cells. Cells were fixed in paraformaldehyde and counterstained with DAPI in order to highlight the nucleus. Secondary antibody: goat anti-rabbit IgG conjugated to Cy3. Please refer to abreview for further details.
ab12147 staining in human testis, showing staining of the cells in the ductus seminiferus (in brown). Paraffin embedded testis tissue was incubated with ab12147 (1:175 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab12147 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
Western blot - Geminin antibody (ab12147)
Lane 1 : Anti-Geminin antibody (ab12147) at 1 µg/ml Lane 2 : Anti-6X His tag® antibody - ChIP Grade (ab9108) at 1 µg/ml
Lane 1 : Recombinant Human Geminin protein (ab40570) Lane 2 : Recombinant Human Geminin protein (ab40570)
Lysates/proteins at 0.1 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Liu P et al. Cell-cycle-regulated activation of Akt kinase by phosphorylation at its carboxyl terminus. Nature508:541-5 (2014).
Read more (PubMed: 24670654) »
Maya-Mendoza A et al. Immortalised breast epithelia survive prolonged DNA replication stress and return to cycle from a senescent-like state. Cell Death Dis5:e1351 (2014).
Read more (PubMed: 25058425) »